欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (19): 9-13.doi: 10.11924/j.issn.1000-6850.2013-0473

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

巴西橡胶树水解酶基因克隆与表达分析

高伟 韦家少 何鹏 吴敏 吴炳孙 王大鹏 高乐 覃怀德   

  • 收稿日期:2013-02-25 修回日期:2013-03-20 出版日期:2013-07-05 发布日期:2013-07-05

Cloning and Expression Analysis of Hydrolase Gene from Hevea brasiliensis Muell. Arg.

  • Received:2013-02-25 Revised:2013-03-20 Online:2013-07-05 Published:2013-07-05

PDF (PC)

613

摘要： 磷是植物生长发育所需的大量重要元素，而缺磷是提高橡胶产量的重要限制因子，研究橡胶树耐低磷胁迫的分子机制，对提高橡胶树磷养分的利用效率具有重要意义。利用低磷胁迫差减文库获得的EST，通过RT-PCR和RACE技术克隆橡胶树应答低磷胁迫基因，并利用实时荧光定量PCR技术进行表达分析。结果表明，从橡胶树中克隆到1个应答低磷胁迫的水解酶基因Hbhyd，全长1860 bp，包含1个1479 bp的完整阅读编码框(ORF)，编码56.1 kD(492aa)蛋白，经NCBI同源性分析，HbHyd编码的氨基酸序列与蓖麻水解酶的氨基酸序列一致性为84%。利用荧光定量PCR分析Hbhyd的表达水平，可知在低磷磷胁迫处理下，HbHyd在橡胶树根中表达量明显上调。Hbhyd基因在橡胶树应答低磷胁迫过程中上调表达，该基因的克隆有助于提高橡胶树磷素吸收利用效率。

关键词: 青贮品质, 青贮品质

Abstract: Phosphorus (P) is an essential macronutrient for plant growth and development. Phosphorus deficiency is a limiting factor for the rubber yield, and understanding molecular mechanism of rubber tree in response to phosphorus deficiency will be helpful to improve phosphorus use efficiency. According to EST from subtractive libraries for roots of rubber tree growing under phosphorus deficient conditions, the gene was cloned by RACE-PCR method and expression analysis was performed by quantitative realtime PCR. The author cloned a HbHyd gene in responding to phosphorus deficiency. Full length of HbHyd was 1860 bp, contained one ORF (open reading frame) with 1479 bp. The NCBI homology analysis showed that the deduced amino acid sequence of HbHyd was 84% identical to a castor (Ricinus communis) hydrolytic enzyme. The results showed that: HbHyd was up-regulated under phosphorus deficiency in the rubber tree roots. HbHyd was up-regulated in responding to phosphorus deficiency in the rubber tree. HbHyd would be helpful to improve phosphorus use efficiency.